HANTAVIRUS VACCINE

"Development of a recombinant vaccine candidate against Hantavirus based on the surface glycoproteins of the Andes virus"

Hantaviruses are viruses transmitted mainly by rodents, capable of generating diseases in humans. According to their geographical location and pathology they are classified into two large groups: the so-called Hantavirus of the old world, present mainly in Europe and Asia, which cause Hemorrhagic Fever with Renal Syndrome (FHSR), and those of the New World, present in America and that cause Hantavirus Cardiopulmonary Syndrome (SCPH).

The Andes virus is the main hantavirus of South America and is the cause of SCPH in Chile. SCPH is a serious disease, which can be deadly. In our country, more than one thousand cases have been confirmed, with mortality between 30% and 40%. A particular feature of the Andes virus is its ability to be transmitted from person to person, unlike other hantaviruses that are transmitted mainly through aerosols of excreta from infected rodents.

Currently, there are no vaccines or treatments against SCPH. Only palliative treatment, which includes circulatory and respiratory supportive therapy in the Critical Care Unit (UPC), may be given to infected patients and may require the use of the Extracorporeal Oxygenation Membrane (ECMO). This is compounded by the difficulty of diagnosing the disease since most of its initial symptoms are similar to flu or cold, and its rapid progression, which leads to pulmonary and cardiac involvement of the patient.

Almost since its creation, the Center for Biotechnology and Spa Biomedicine. has been working on the development of an effective vaccine against hantavirus. The vaccine is composed of the ectodomains of the Gn and Gc glycoproteins of the Andes virus. In nature, both molecules are exposed on the surface of hantaviruses, and are the main target of the immune response when a person is infected with the pathogen. The Center for Biotechnology and Spa Biomedicine. has managed to establish up-and downstream processes for the production, by recombinant route, of both antigens. The recombinant molecules have been characterized both biochemically and antigenically. The immunogenicity of these molecules has also been demonstrated in murine and Syrian hamster models. The project is currently focused on the production of the vaccine candidate under GMP conditions, for its preclinical and clinical phase 1 validation.

This project is co-financed by Corporación de Fomento de la Producción (CORFO), under the code 17IDAE-74735.

ANTIBODIES IN GOAT MILK

"Development and characterization of biobetter variant of human erythropoietin"

Cancer is one of the leading causes of morbidity and mortality worldwide; in 2012 there were 14 million new cases and 8.2 million deaths related to cancer. In 2012, the cancers that caused the highest level of mortality were those of the lung (1.59 million deaths), liver (745,000 deaths), gastric (723,000 deaths), colorectal (694,000 deaths), breast (521,000 deaths) and esophagus (400,000 deaths). It is anticipated that annual cancer cases will increase from 14 million in 2012 to 22 million in the next two decades. Despite conventional treatments of chemo and radiotherapy, the average 5-year survival rates are considerably low: lung cancer (15%), liver cancer (12%), stomach cancer (26%), colorectal cancer ( 66%), breast cancer (77%), esophageal cancer (21%). Monoclonal antibodies are currently used for cancer treatment, since they have proven to be a more specific, less toxic therapeutic alternative, and consequently more effective in terms of their capacity to increase patients' time and quality of life, as well as the total survival rate. However, the production of these drugs is carried out in cultures of mammalian cells. This significantly increases the production costs, directly affecting the patient, since they are expensive treatments impossible to finance by patients or by any public entity in the country.

That is why the Center for Biotechnology and Spa Biomedicine. works in the production of antitumor antibodies in the milk of genetically modified goats. This modification is localized, that is, only the mammary gland will be genetically modified so that anti-tumor antibodies are produced in the milk of the goat. This does not report any type of alteration or suffering in the animal. By using the mammary gland as a bioreactor, we will obtain antibodies at a lower production cost. Currently working on the effectiveness tests of the new antitumor antibodies.

This project is co-financed by Corporación de Fomento de la Producción (CORFO), under the code 16IDAE-67701.

VARIANT BIOBETTER OF HUMAN ERYTHROPOIETIN

"Development and characterization of biobetter variant of human erythropoietin"

Human erythropoietin (hEPO) is the main hormone responsible for stimulating the generation and differentiation of red blood cells, therefore, it is essential for the oxygenation and renewal of blood. This protein is produced in the kidneys and its levels are increased in athletes, people who live or visit high altitude places, divers, etc. The deficiency of hEPO is serious and generates a severe anemia common in patients with chronic renal failure, kidney failure associated with trauma and kidney transplants. This anemia is also associated with the effects of chemotherapy in cancer patients, azidovidine treatments in patients infected with the human immunodeficiency virus (HIV) and patients with diabetic nephropathy. According to the last national hemodialysis account of the Chilean Society of Nephrology (August, 2015) in Chile there are 19,071 people, children and adults in hemodialysis with a rate of 1060 patients per million population (pmp) while in the USA the prevalence of IRCT (Chronic Terminal Renal Insufficiency) is 1,131 pmp with an incidence of 296 new patients per year.

For the treatment of all these patients, the external and constant supply of recombinant human erythropoietin (rhEPO), produced in the laboratory, is essential. The rhEPO is currently one of the 10 most sold biopharmaceuticals in the world, however, it is still not satisfied to meet the demand due to its high cost. In Chile only 49.4% of patients on Chronic Hemodialysis (HDC) use rhEPO for the maintenance of erythropoiesis, despite the fact that the treatment is part of the AUGE / GES system, this is because the State reserves it only for critical patients. The weekly treatment (3 doses) per patient costs the state between 6,000 and 10,000 pesos, while in a particular way it can cost between 60,000 and 100,000 pesos depending on the variant used. Since 2010, the Chilean government invests more than 2000 million Chilean pesos annually in the purchase of this drug.

The Center for Biotechnology and Spa Biomedicine. works in the generation and evaluation in vitro and in vivo of a new, improved variant of rhEPO. This new molecule can be produced at a lower cost than the current one and we estimate that it will have a half-life in blood 60 times higher than the conventional rhEPO. In this way, the number of doses needed for the patient and the total cost of treatment could be reduced.

This project is co-financed by Corporación de Fomento de la Producción (Corfo), under the code 16IDAE-67704.

VARIANT BIOBETTER OF FSH BOVINE

"Enabling capacities to develop an innovative product for the regulation of heat and pregnancy"

The growing global need for food for human consumption raises the need to produce larger volumes of food, with a high nutritional value and in an increasingly efficient way. In this context, the livestock industry in order to increase its productive efficiency has been forced to select individuals with increasingly better genetic and phenotypic characteristics, such as resistance to a particular climate, higher feed conversion rates, greater reproductive capacity, etc. Conventional or natural fertilization is limited by the number of offspring that a female is capable of producing, which under natural conditions does not exceed one calf per year in the case of bovines, and therefore, the 8 calves throughout their lives reproductive The assisted fertilization, in this sense, manages to improve the reproductive capacity of the stock farms and ensures that these pregnancies come to term in a correct way, thus speeding up the selection process compared to conventional breeding.

Currently there are a number of technological tools to facilitate the reproductive process in livestock such as heat synchronization, in-vitro fertilization, artificial insemination and embryo transfer. Much of these technical procedures are performed during the assisted fertilization protocols and require the exogenous administration of hormones such as progesterone, prostaglandins, follicle stimulating hormone (FSH) and luteinizing hormone (LH), among others, which allow to emulate and take advantage of reproductive processes natural in order to increase reproductive efficiency.

The hormones available in the current market are obtained almost entirely from pituitary gland extracts from slaughtered pigs in slaughterhouses, and although they are widely used, important limitations have been reported that affect the certification of production processes, implying high risks of contamination with pathogens, contamination with other hormones that interfere in different degrees with the processes of assisted fertilization and important variations of the active ingredients between each production lot.

The company Biotechnology and Biomedicine Center has obtained a Biobetter variant of FSH, indicated for use in heat synchronization protocols and stimulation of superovulation in cows and heifers suitable for reproduction. This artificial variant of the bovine FSH hormone, developed under the strictest quality standards, has been created specifically to solve many of the problems presented by FSH obtained from pituitary pigs. In this context, the structural modifications incorporated in the molecule allow it to maintain its bioactivity in circulation for much longer, thus simplifying the administration protocols and decreasing the daily doses that must be applied. In addition, thanks to its recombinant origin, the risks of contamination with pathogens and other hormones are eliminated, it allows scaling of the production process and ensures homogeneity between batches, which favors the obtaining of reproducible results.

This initiative was initially funded by the Corfo Prototype project of Regional Innovation Code 16PIRE-60724 between the period of July 2016 to June 2017, which allowed obtaining a prototype of the solution and winning the project Enabling innovation for the regional SME, code 17 -HIBIO-78583, included between August 2017 to July 2018 to materialize the production of the hormone. Currently the company has developed the capacities to supply commercially to the national market, being available licensing agreements of production technology to interested international companies.

BLOCKING MOLECULES OF TNFalpha

"Chronic inflammatory diseases"

The blocking of tumor necrosis factor alpha (TNFα) is a therapeutic alternative for the treatment of multiple chronic inflammatory pathologies, among them: rheumatoid arthritis, juvenile idiopathic arthritis, ankylosing spondylitis, psoriatic arthritis, ulcerative colitis, and Crohn's disease. Currently, there are 5 antibodies on the market, or fragments of antibodies, capable of blocking TNFα. Due to its therapeutic efficacy, and a large number of pathologies that require its use, these 5 molecules have become the best selling biopharmaceuticals in the world, with global sales that, in 2016, reached 36,000 million dollars. Despite its therapeutic efficacy, anti-TNFα biopharmaceuticals are economically inaccessible for most patients require it. In Chile, the monthly cost of treatment with TNFα blockers ranges between 700,000 and 900,000 pesos.

With this background, the Center for Biotechnology and Spa Biomedicine. works in the development of a new generation of TNFα blockers based on small peptides. Compared with therapeutic antibodies, the peptides can be produced by chemical synthesis, the preparations more homogeneous, are more stable, and their production costs are significantly lower. The sum of these advantages should be translated into a new type of TNFα blocking drug with a sale price between 3 and 6 times lower than the current biopharmaceuticals.

In this line of research, the company has already identified peptides with proven ability to block TNFα. The current works are aimed at increasing the average lifetime in circulation of these molecules, in order to enhance their therapeutic effect.

This project is co-financed by Corporación de Fomento de la Producción (Corfo), under the code 16IDAE-67705.

DIABETIC FOOT TREATMENT

"Therapeutic formulation for tissue regeneration in patients with diabetic foot ulcers"

In Chile, more than 9.4% of the population is affected by Diabetes mellitus, exceeding the global average for this pathology by more than three times. It is estimated that 12.5% of the Chilean diabetic population develops the pathology called Diabetic Foot Ulcer (UPD), of which approximately 12% must undergo amputation of their limbs after the injuries worsen. The UPD is characterized by a loss of sensibility of the foot or leg that allows any cut or trauma to go completely unnoticed for days or weeks, this added to the low cell regeneration product of the decreased blood supply in the area facilitates the progression of the wound and the possibility of infection.

The Chilean health system does not have effective drugs that minimize or cancel the cellular damage produced during the progression of the ulcer, which makes it difficult to access complementary care alternatives for affected patients. On the other hand, the effective alternatives available need to be imported from abroad at a cost per treatment that in some cases exceeds USD $ 6,000 per patient, making them virtually inaccessible to the bulk of the affected population.

Recently, the Center for Biotechnology and Biomedicine has generated a topical application formulation made from the natural polymer Chitosan that acts as a structural component and regenerative adjuvant, and recombinant human Epidermal Growth Factor as an Active Pharmaceutical Ingredient (IFA), which acts by stimulating the repair of damaged tissue. This IFA is produced through an innovative process of production of a recombinant base that uses the yeast Pichia pastoris as an expression system, and a series of steps of chromatographic purification that allows to obtain it with a very high level of yield and purity. Currently, this technology is in a state of production established at the laboratory level and with its biological function validated in two animal models, pigs, and sheep.

This Initiative was initially funded by the Innova Applied R & D Project, Code 13IDL2-23507, which ran from the December 2013 period until June 2016 and later by the Innovations Validation and Packaging project, code 17ITE2-78771, included in the period of September 2017 to August 2019.

GLYCOSYLATION PATTERNS IN MAMMARY GLAND

"Modification of the mammary epithelial cell glycosylation pathway for the efficient production of recombinant proteins"

Currently, there is a growing over-demand of protein biopharmaceuticals because the needs of the population exceed the productive capacities of the pharmaceutical industry worldwide. The proteins that are used as biopharmaceuticals have a high degree of structural complexity and their active form can only be expressed in artificial cultures of cells from human or mammalian tissues, since they need to go through a specific glycosylation process that only occurs naturally in cells mentioned above, and whose function is to ensure the correct folding of the protein, ultimately influencing its functionality.

The main complex biopharmaceuticals in high demand at present are human erythropoietin, insulin, growth hormone, and interferon alpha / beta. These are generally used in the treatment of chronic diseases associated with the deficient production of these proteins, however, their expression in artificial cellular systems represents a high production cost.

In this sense, the use of animals as bioreactors appears as an efficient alternative to the production of biopharmaceuticals in cell cultures in vitro, since the costs of production and maintenance of the latter are considerably higher compared to production in in vivo systems. The Center for Biotechnology and Biomedicine has been able to produce the hormone erythropoietin (EPO) in mammary gland epithelial cells, carrying one of the best-selling biopharmaceuticals in the world for the treatment of anemia in patients with chronic renal insufficiency, immunosuppressed and in treatment against cancer towards an in vivo production. The EPO produced also has a glycosylation pattern of tetra-antennary type and finished in sialic acid, which ensures that the molecule is biologically active. This was achieved through the overexpression of key enzymes of the glycosylation pathway such as GnTIV in conjunction with EPO in mammary gland cells, representing an important advance for the center and positioning it as one of the pioneering groups in the country to achieve the modification of the glycosylation pathway in the mammary gland of goats. This allows us to project the work done towards obtaining other proteins of therapeutic interest that require this glycosylation pathway to reach their biological activity, which directly impacts the scaling capacity and differentiation of the technology.

This Initiative is financed by the Innova Applied R & D Project, Code 17-IDAE-74738, comprised between the period of September 2017 and August 2019.

ALTERNATIVE TO SURGICAL CASTRATION IN MALE BOVINE

"Prototype for the castration of males in the bovine sector"

In cattle farming, every day becomes more important to avoid practices that may cause pain or damage to animals. One of the main activities affecting animal welfare in the bovine sector is the castration of males, which is used with the aim of having more docile animals for handling and obtaining a better quality of meat, since castrating increases fat coverage, chromatic pigmentation, and tenderness of the meat. At present 90% of calves in Chile are castrated, however, some side effects have been observed such as the physiological stress that is generated after the procedure, the reduction of the plasma concentration of anabolic hormones and the reduction of the gain of Daily weight that can reach up to -15% per day. To counteract the effects on the loss of meat production, the livestock industry has implemented the use of anabolics using subcutaneous implants, which are widely used to promote the production of animal protein in the various farms in the country. However, its use has limited exports to countries such as China and the European Community, which prohibit the administration of anabolic substances due to the harmful side effects that their residues would cause in meats destined for human consumption. Considering this, our country to maintain its position in the meat export market, must comply with the requirements of these in terms of animal welfare, as well as the elimination of the use of anabolic compounds in their products, so there is a need to to be able to count on systems that allow castrating the animals with less pain and that also do not entail a productive loss, in order to be able to avoid the use of anabolics.

Given this opportunity, the Center for Biotechnology and Biomedicine has developed a solution for the castration of males through a sclerosing formulation as an alternative to traditional castration methods. This formulation eliminates the need to establish a surgery for the procedure and does not completely eliminate the testosterone-producing cells, so it does not affect the daily weight gain, but behavioral behavior similar to that of an animal castrated by traditional methods is achieved. like surgical castration.

This Initiative is financed by the Regional Innovation Prototype Project code 17-PIRE-72263, between the period of May 2017 to April 2018.

IMMUNOMODULATOR FOR USE IN SWINE

"Development of a nanoencapsulated prolonged release formulation with therapeutic and / or prophylactic effects based on recombinant porcine interferon alpha for porcine veterinary applications"

Pork meat is one of the most consumed worldwide with about 36% consumption compared to other alternatives, a demand that increases constantly as the population grows. This growth, however, has not gone hand in hand with production figures, which has even gone down showing an annual decrease of between 8 to 10% due to the closure of some hatcheries product of outbreaks of infectious diseases. To supply the shortage, hatcheries have intensified the production process by decreasing the suckling time of the piglets to put them quickly into fattening. This has meant important problems associated with the affective, alimentary and immune depression that causes weaning in pigs, opening the door to opportunistic infections that affect the normal development of piglets. In this context, the release of certain immune proteins has been observed in the face of viral infections, which are known as interferons. These interferons are characterized by having antiviral action and act as modulators of immune response, so it is suggested that their role as reinforcers of the immune system would be key to prophylactically control viral infections. To date, there is no alternative available in the market in pigs in the weaning stage that allows generating an early maturation of the immune system and protection against viral pathologies other than antibiotics, probiotics or antiviral vaccines. This is why the idea of developing a sustained release system that contains interferon alfa would strengthen the immune system throughout the weaning period, allowing the animals to respond adequately to any type of viral infection.

In this sense, the Center for Biotechnology and Biomedicine in conjunction with the Veterinary Company, have worked together to develop a nanoencapsulated solution based on porcine interferon alfa obtained by recombinant, which will be used by the veterinary market for use as immunomodulator in pigs and as a coadjuvant in various vaccine formulations applied during the production process. This solution aims to reduce the losses associated with infectious diseases in the pig sector, becoming an innovative, effective and low-cost alternative.

This initiative is financed by the I + D Applied to the Business project, code 17-IDAE-74707 during the period from August 2017 to July 2019.

GROWTH STIMULATORS FOR SALMONIDS

"Validation of a peptide formulation to stimulate growth and resistance to pathogens in salmonids"

Due to the high demand presented by salmonids today, the aquaculture industry has been forced to develop intensive farming conditions at high population densities to meet the high demand and growth levels required by the industry and the market. These culture conditions favor the presence of stress factors to which the fish are subjected, which are also increased by the multiple processes of vaccination, transport and cleaning to which they are subjected. This constant condition of stress in fish affects, among other problems, the reduction of growth, the deterioration of the immune response and, therefore, the increase in susceptibility to infectious agents. The economic losses in the industry associated with this concept are considerable, so the optimization of cultivation strategies that involve strengthening the immune response to avoid the susceptibility to contracting diseases during the production process are key to the industry.

Conventional treatments are based on the use of antibiotics and antivirals to reduce the incidence of these pathogens, which results in a problem of resistant microorganisms, the appearance of antibiotic residues in fish and environmental damage.

Based on this problem, the Center for Biotechnology and Biomedicine has developed a formulation based on synthetic peptides that stimulate growth, survival and the immune system of fish, applicable at different stages of the salmonid culture cycle. This formulation has as main attributes the increase in the growth rate, the reduction of the feed conversion factor, stimulation of the innate immune system and the enhancement of the acquired immune system, which contributes to the decrease in the use of antibiotics with the consequent impact positive in production costs and in the environment.

This initiative is funded by the Corfo Validation and Packaging of Innovations project, Code N ° 16ITE2-60263 between August 2016 until July 2018.